The present invention relates to the identification of a novel use of the principles extracted from the plant Murraya koenigii for treatment of asthma. This invention provides a process for the extraction of the principles from the plant Murraya koenigii. The invention also provides novel compositions useful in the treatment of asthma and methods for the treatment of asthmatic conditions using the said composition.
Respiratory diseases such as asthma are reaching epidemic proportions in both the developed and developing world (Nature, Vol. 402, Supplement, and No. 6760, 1999. A special supplement on allergy and asthma). In countries such as Britain and Australia, the respiratory problem translates to 1 in 4 children under age of 14 years having asthma. The disease causes distress and misery in millions often at a time in their lives when they should be most active. Asthma interferes with sleep, intellectual functioning and recreational activities. At one extreme, asthma can be life threatening leading to occurrence to deaths that are avoidable.
Expert panel report has offered guidelines for diagnosis and management of asthma (JAMA Asthma Information Center, www.amaasssm.org/special/asthma/treatmnt/guide/guideline/gudelin.htm). Control of factors contributing to asthma severity is another avenue for management (JAMA Asthma Information Center, www.amaasssm.org/special/asthma/treatmnt/guide/guidelin/comp2/comp2.htm). several therapies and medications have also been suggested (JAMA Asthma Information Center, www.amaasssm.org/special/asthma/treatmnt/guide/guidelin/comp3/medicat/meditoc.htm and www.amaass.org/special/asthma/treatmnt/guide/guidelin/comp3/comp3.htm). Immunotherapy is another route for the treatment (JAMA Asthma Information Center, www.amasssm. or/special/asthma/treatmnt/guide/guidelin/comp2/immunoth.htm). Complementary alternative medicine is also being tried (JAMA Asthma Information Center, www.amaasssm.org/special/asthma/treatmnt/guide/guidelin/comp3/medicat/alternat.hmt) However, no satisfactory medication for cure of the disease has been found.
Herein the active factors in Murraya koenigii was prepared and used under in vitro system to show its usefulness for relief, treatment and cure of asthmatic problem. The plant Murraya koenigii has been studied and reported to have various medicinal values (Chakraborty, M. P Phytochemistry 1997, October; Khan B. A, Indian J of Expt. Biol. 1997 February; Khan B A Indian J of Physiology and Pharmacology 1996, April; Khan B A Plant Foods Hum. Nutr. 1996 June).
The leaf of this plant is widely used in various food preparations in India. Further, the plant is ubiquitous. Thus the prior art suggests that the extracts of the plant have several medicinal and other properties which have been widely investigated. This is adequate proof that the extracts of the plant can be safely consumed by humans without any side effects.
The inventors undertook a study on the extracts of this plant to identify the therapeutic principles of the plant. During their study, the inventories discovered the presence of the active principles in the leaf of Murraya koenigii which surprisingly were found to be useful in the treatment and cure of asthma.
The main objective of the present invention is to provide a method for the treatment of asthmatic conditions employing active principles extracted from the plant Murraya koenigii. 
Another object is to provide a process for the isolation of active principles from the plant Murraya koenigii and use the same for relief, treatment and cure of asthma.
Yet another object is to provide a simple, fast and inexpensive process to obtain a mixture of active (four) compounds possessing biological activities useful in the treatment of asthma.
Still another object is to provide novel compositions containing active principles extracted from the plant Murraya koenigii and useful in the treatment of asthma.
Accordingly, the present invention provides a method for the treatment of asthma, employing principles extracted from the plant Murraya koenigii. The invention also provides novel compositions useful in the treatment of asthma and methods for the extraction of active principles from the plant Murraya koenigii. 
Accordingly, the invention provides a process for preparing an extract from the plant Murraya koenigii, useful in the treatment of asthma, said process comprising the steps of pulverising plant materials obtained from plant Murraya koenigii, extracting the plant material with a solvent at ambient temperature, concentrating the extract by filtering and evaporating it under reduced pressure and lyophilizing the concentrate to obtain a lyophilized extract containing active principles of the plant Murraya koenigii. 
In an embodiment, the plant materials are obtained from plant parts of Murraya koenigii selected from garden fresh leaves or leaves dried under shade.
In another embodiment, the leaves are pulverized by conventional methods to get homogenized leaves.
In yet another embodiment, the plant materials are extracted with solvents selected from hydrocarbon solvents, chlorinated solvents, ester solvents, ketonic solvents, alcohols, water and buffers.
In still another embodiment, the solvents are selected from the group consisting of petroleum ether (BP 40-60xc2x0 C.), petroleum ether (60xc2x0 C.-80xc2x0 C.), benzene, pentane, hexane, chloroform, dichloromethane, carbon tetrachloride, diethyl ether, tetrahydrofuran, dioxane, acetone, cyclopentanone, ethyl acetate, ethyl formate, methanol, ethanol, n-butanol, water and buffers. In another embodiment, the concentration of the extract is effected by filtering and evaporating the solvents under reduced pressure at a temperature range of 20xc2x0 C.-80xc2x0 C. preferably at ambient temperature and lyophilizing the concentrate by conventional methods to obtain mixtures of the active factors.
In yet another embodiment, the extract obtained from the plant Murraya koenigii comprises active principles which dark colored solids soluble in dimethylsulfoxide.
In another embodiment, the active principles obtained from the plant Murraya koenigii are biocompatible and non-toxic in nature. The active principles have Rf values 0.73, 0.60, 034 and 0.14 in chloroform and methanol in the ratio 19:1 and Rf values 0.60, 0.38, 024 and 0.15 in the chloroform. The active principles have four peak with retention time 3.37, 3.49, 4.0 and 5.69 in methanol as solvent at 254 nm. The extraction process is carried out for a period ranging from 1-120 hrs, preferably between 12-16 hrs.
Further, the invention also provides pharmaceutical composition useful in the treatment of asthma, said composition comprising an effective amount of extract obtained from the plant Murraya koenigii together with, or optionally associated with a pharmaceutically acceptable additive. The additives used in the composition comprise powder or extracts of plants selected from M. panicitlate Linn, H. abelmoschus, T ammi, S. aromaticum, A. vasica Nees, E hirta, and M. koinegii. These additives are present in the range of 80-100 mg of M. paniculate Linn, 40-60 mg of H. abelmoschus, 38-62 mg of T. ammi, 7-13 mg of S. aromaticum, 85-115 mg of A. vasica Nees and 90-110 mg of E hirta. 
In an embodiment, the composition comprises
In the said composition, the extract of the plant M. koiitegii is present in the range of 87-105 mg per dose. In the said composition, the extract of the plant M. koinegii comprises active principles which are dark colored solids, soluble in dimethylsulfoxide. The active principles have Rf values 0.73, 0.60, 034 and 0.14 in chloroform and methanol in the ratio 19:1 and Rf values 0.60, 0.38, 024 and 0. 15 in the chloroform. These principles have four peaks with retention time 3.37, 3.49, 4.0 and 5.69 in methanol as solvent at 254 nm. The said active principles obtained from the plant M. koinegii exhibit antioxidant property i.e. O2 inhibition.
Further, the invention provides a method for the treatment of asthma, said method comprising the steps of administering an effective amount of the composition comprising an extract obtained from the plant Murraya koenigii and at least one pharmaceutically acceptable additive to a subject in need thereof.
In an embodiment, the lyophilized extract obtained from Murraya Koenigii is administered alone or along with other conventional additives for the treatment of asthma. In still another embodiment, the mode of administration is oral for the treatment of mild or acute asthma.
In yet another embodiment, the dosage level of the composition, comprising the extract from the plant Murraya koenigii is between 325-600 mg twice daily for the period ranging from 3 to 30 days.
In another embodiment, the dosage level is in between 325-600 mg twice daily for the period ranging from 3 to 15 days for mild asthmatic condition, and 15-30 days for acute asthmatic condition.
In still another embodiment, the additives are selected from M. panicutlate Linn, H. abelmoschus, T. ammi, S. aromaticum, A. vasica Nees, E hirta, and M. koinegii. 
As said above, the additives are present in a range of 80-100 mg of M. paniculate Linn, 40-60 mg of H. abelmoschus, 38-62 mg of T. ammi, 7-13 mg of S. aromaticum, 85-115 mg of A. vasica Nees, 90-110 mg off E. hirta, and 87-105 mg of M. koinegii. per dose.
In another embodiment, the additives obatined from M. paticulate Linn, H. abelmoschus, T. ammi, S. aromaticum A. vasica Nees, E. hirta, and M. koinegii are administered to include properties such as antidiahorial, antiseptic, carminative, stimulation, anti-cough, anti-bronchitis and nourishment.
In another embodiment, the additives obtained from M. paniculate Linn, H. abelmoschus, T. ammi, S. aromaticum, A. vasica Nees, E hirta, and M. koinegii are administered to include properties such as antidiarrheal, antiseptic, carminative, stimulation, anti-cough, anti-bronchitis and nourishment.
In addition, the invention provides an anti-oxidant composition for human beings and animals, said composition comprising a effective amount of extract obtained from the plant Murraya koenigii together with or optionally, associated with pharmaceutically acceptable additives.
In still another embodiment, the additives comprise powder or extracts of plants selected from M. pamiclate Linn, H. abelmoschus, T. ammi, S. aromaticum, A. vasica Nees, E hirta, and M. koinegii. 
In an embodiment, the additives are present in a range of 80-100 mg of M. paniculate Linn, 40-60 mg of H. abelmoschus, 38-62 mg of T. ammi, 7-13 mg of S. aromaticum, 85-115 mg of A. vasica Nees, 90-110 mg of E hirta, and 87-105 mg of M. koinegii. per dose of the anti-oxidant composition.
As said above, the active factor(s) in Murraya koenigii useful for relief, treatment and cure of asthmatic problem(s), the preparation of which comprises drying, powdering, and extracting the dried leaves of the plant, Murraya koenigii, in a percolator at an ambient temperature using appropriate solvents and concentrating the extract under reduced pressure and finally lyophilizing the concentrate to make the active factor(s).
In the present invention, active factor(s) in Murraya koenigii for the treatment and cure of asthmatic problem are prepared from the leaves of the plant by drying, powdering and extracting, in a percolator at an ambient temperature using appropriate solvents and concentrating the extract under reduced pressure and finally lyophilizing the concentrate to make the active factors.
In an embodiment of the present invention, the active factors from the plant material to be used as an oral preparation for relief, treatment and cure of asthmatic problem.
In another embodiment of the present invention, the leaf of Murraya koenigii used was garden fresh or fresh leaf dried under sun or shade.
In still another embodiment of the present invention, plant material was used for the extraction with appropriate solvents such as methanol or water or buffers in a percolator.
In yet another embodiment of the present invention, the plant extract made by the methanol/appropriate solvent or water was concentrated under reduced pressure to get the mixture of active factor.
In yet another embodiment of the present invention, the concentrate of the plant extract was lyophilized to make the factors free of solvent or water.
In yet another embodiment of the present invention, lyophilized solid obtained as the mixture of active factors present in the leaf of Murraya koenigii was proposed to be used for relief, treatment and cure of asthmatic problem.
The method of preparation of the active factors comprises the following
1) collecting the fresh leaves from the local suppliers,
2) drying the leaves under shade to a moderate degree or to take the fresh leave as the starting material,
3) powdering the dried or homogenizing the fresh leaves in the apparatus known in the art,
4) putting the powder or homogenate in a percolator under the bulk of appropriate solvents; choosing hydrocarbon solves such as petroleum ether (B.P 40-60xc2x0 C.), petroleum ether (B.P 60-80xc2x0 C.), pentane, hexane, benzene etc.; chlorinated solvents like chloroform, dichloromethane, carbon tetrachloride etc.; etherial solvents such as diethyl ether, tetrahydrofuran, dioxane etc., ketonic solvents such as acetone, cyclopentanone etc.; ester solvents such as ethyl acetate, ethyl formate etc; all alcohol such as methanol, ethanol, n-butanol etc.; water and buffers,
5) extracting the percolated plant material using a percolator or the apparatus or equipment currently known in the art over a period of time ranging from 1 to 120 hours,
6) evaporating the solvent under reduced pressure using an apparatus or equipment currently known in the art at a temperature ranging from 20 to 80xc2x0 C.,
7) lyophilizing or drying the material in the apparatus or equipment currently known in the art,
8) storing the processed material in a cool and dry place in an air tight container, and
9) evaluating the bioactivity of the material.
The following examples are given by way of illustration and therefore should not be construed to limit the scope of the present invention.
Collection of Plant Material
The leaves of M. koenigii were collected from shrubs in the month of March-August 2000 from different areas of West Bengal, India. A voucher specimen is deposited at the department of Medicinal Chemistry at the Indian Institute of Chemical Biology, 4, Raja S. C. Mallick Road, Calcutta-700 032.
Preparation of the Active Factors or Material